USPTO Patents Application 09544084

PCX

WORLD INTELLECTUAL PROPERTY ORGANIZATION
lnt£maiionat Burrau

INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)

(51) International Patent Classification ^
A61K 31/20

Al

(lU International Publication Number: WO 97/46230

(43) International Publication Dale: II December 1997 (11.12.97)

(21) International Application Number: PCT/L;S97/04538

(22) International Filing Date: 19 March 1997 (19.03.97)

(30) Priority Data:

08/659.845
08/736,562

7 June 1996 (07.06.96) . US

28 August 1996 (28.08.96) US

(71) Applicant: WISCONSIN ALUMNI RESEARCH FOUNDA-

TION [US/USJ; P.O. Box 7365. Madison, Wl 53707-7365
(US).

(72) Inventors: COOK. Mark, E.; 15 Kewaunee Coun, Madison,

W! 53705 (US). PARIZA, Michael. W.; 7102 Valhalla
Trail. Madison. Wl 53719 (US).

(74) Agent: KRYSHAK. Thad; Quarles & Brady. 41 1 East Wiscon-
sin Avenue. Milwaukee, Wl 53202-4497 (US).

(81) Designated Slates: AL. AM. AT. AU, AZ. BA, BB, BG. BR.
BY. CA. CH. CN. CU. CrZ. DE. DK. EE. ES. Fl. GB. CE.
GH. HU. IL, IS, JP. KE, KG. KP. KR. K2. LC uK. LR.
LS. LT. LU. LV. MD. MG. MK. MN, MAV. MX. NO. N2.
PL. PT. RO. RU. SD. SE. SG. SI. SK. TJ. TM. TR. TT.
UA, UG. UZ, VN. YU. ARFPO patent (GH. KE. LS. MW.
SD. SZ. UG). Eurasian patent (AM. AZ. BY. KG. KZ. MD.
RU. TJ. TM). European paicnt (AT. BE. CH. DE, DK. ES.
FI. FR. GB. GR. IE. IT. LU. MC, NL. PT. SE). OAPI patent
(BF. BJ. CF. CG. CI. CM. GA. GN. ML. MR. NE. SN. TD,
TG).

Published

With international search report.

(54) Title: METHOD FOR MAINTAINING AN EXISTING LEVEL OF BODY FAT AND/OR BODY WEIGHT
(57) Abstract

A method of maintaining an existing level of body fat or Ixxly weight in a human which comprises administering to a human desiring
to maintain that enlisting level a safe and effective amount of conjugated linoleic acid (CLA) to maintain that level.

FOR THE PURPOSES OF INFORMATION ONLY

Codes used to identify Slates party to the PCX on the front pages of pamphlets publishing international applications under the PCX.

AL

AlbMU

ES

Spain

LS

l.cso(ho

SI

Slovenia

AM

Armcnit

Fl

FinlMd

LT

Lithuania

SK

Slovakia

AT

Auttria

FK

France

LU

Luirmbcnirg

SN

Senegal

AU

Australia

GA

Gabon

LV

Latvia

sz

Swaziland

AZ

Aserfatijan

CB

United Kingdoin

MC

MoAaco

TO

Chad

BA

Bosnia and Herzegovina

GE

Georgia

Ml)

Republic of Moldova

TC

Togo

BB

BartMdot

GH

Ghana

MG

Madagascar

TJ

Tajik iuan

BE

Belgium

GN

Guinea

MK

The former Yugoabv

TM

Turkmenutan

BF

Bufttna Faso

GR

Greece

Republic of Macedonia

TR

Turkey

BG

Bulgaria

HU

Hvmgary

ML

Mali

XT

Trinidad and Tobago

BJ

Benin

IE

trelvid

MN

Mongolia

UA

Ukraine

BR

Brazil

IL

timet

MR

Mauriunia

UG

Uganda

BY

Belarus

IS

Iceland

MW

Malawi

US

United Slates of America

CA

Canada

IT

haly

MX

Meaico

uz

Uzbekistan

CF

Central African Republic

JP

lapwi

NE

Niger

VN

Vtef Nam

CG

Congo

KE

Kenya

NL

Nethertandt

YD

Yugoslavia

CH

Swiizeriand

KG

Kyrgywan

NO

Norway

zw

Zimbabwe

a

Cdte d'lvotre

KP

DeRXKTaiic Pieople't

NZ

New Zealand

CM

Cameroon

Republic of Korea

PL

Poland

CN

China

KR

Republic of Korea

PT

Portugal

CO

Cuba

KZ

Kauksun

RO

Romania

cz

Ctech Republic

LC

Saint l^ia

RU

Ruuian Federation

DE

Germany

U

Liectuenitein

SO

Sudan

DK

Oenmark

l,K

Sri LvAa

SE

Sweden

EE

LR

Libertt

SC

Singapore

wo 97/46230

PCT/US97/04538

-1-

METHOD FOR MAINTAINING
AN EXISTING LEVEL OF BODY FAT AND/OR BODY WEIGHT

Field of the Invention

The present invention generally relates to human
nutrition. More particularly, it relates to a method of
treating humans to maintain an existing level of body fat
5 and/or body weight.

Background of the Invention

There are a significant nurrUDer of people, who are happy
with their existing body weights and levels of body fat, but
who do not want their weight or levels of body fat to
increase. In addition, thousands of people annually go on
diets to lose body fat or weight. Unfortunately, most of
those that are successful cannot maintain. the lower levels of
body fat and/or body weight which they have achieved.

There is a need for a method of maintaining an existing
level of body fat and/or body weight in a human.

Brief Summary of the Invention
It is an object of the present invention to disclose a
method of maintaining an existing level of body fat and/or
body weight in a human.
20 We have discovered that an existing level of body fat

and/or body weight in a human can be maintained by
administering to the human a safe and effective amount of an
active form of a conjugated linoleic acid, such as 9,11-
octadecadienoic acid and 10 , l2-octadecadienoic acid, an ester
25 thereof, a non- toxic salt thereof, and mixtures thereof.

The terms "conjugated linoleic acids" and "CLA" as used
herein are intended to include 9, 11 -octadecadienoic acid,
10, 12-octadecadienoic acid and their active derivatives, such
as non- toxic salts and esters, and mixtures thereof.
30 The present method can be used by ex-dieters to maintain

the lower level of body fat and/or body weight they have
achieved or by persons who wish to increase their fat intake
without increasing their level of body fat and/or body weight.

10

15

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-2-

It will be apparent to those skilled in the art that the
f orement ioned objects and other advantages may be achieved by
the practice of the present invention.

Description of the Preferred Embodiment

5 The present method comprises administering to the human

desirous of maintaining his or her existing level of body fat
and/or body weight a safe and effective amount of an active
form of a conjugated linoleic acid, which is selected from a
conjugated linoleic acid, such as 9 , 11 -octadecadienoic acid

10 and 10, 12-octadecadienoic acid, an ester thereof, a non-toxic
salt thereof, and mixtures thereof.

The existing level of the human's body fat and/or body
weight can be determined by a variety of methods. The body
weight of a person can be obtained simply by weighing the

15 person. One method of determining body fat. simply comprises
doing a "pinch test" at the waist, chest, thighs and other
body parts. Another more sophisticated method, which is
commonly used for athletes, involves completely submerging the
person in liquid and calculating the body weight under water.

20 The amount of the CLA to be administered normally is an

amount which is equal to about 1% to about 30% of the fat in
the human's diet. If the CLA is taken in pharmaceutical
dosage form the dose will normally be about 100 mg . to 20,000
mg. per day of CLA in the form of the free acids. Since the

25 CLA is a natural food ingredient and relatively non- toxic, the
amount which can be consumed is not critical as long as it is
enough to be effective and it is not contraindicated because
of the human's health.

The practice of the present invention is further

30 illustrated by the examples which follow:

Example 1

SYNTHESIS OF CONJUGATED LINOLEIC ACIDS (CLA)
FROM LINOLEIC ACID AND SAFFLOWER OIL

35 Propylene glycol (1000 g) and 500 g potassium hydroxide

(KOH) are put into a 4 -neck round bottom flask (5000 ml) . The

flask is equipped with a mechanical stirrer, a thermometer, a

wo 97/46230

PCT/US97/04538

reflux condenser, and a nitrogen inlet. (The nitrogen
introduced in first run through two oxygen traps) .

Nitrogen is bubbled into the propylene glycol and KOH
mixture for 20 minutes and the temperature is then raised to
5 180«C.

1000 g of linoleic acid, corn oil, or saf flower oil is
then introduced into the flask. The mixture is heated az 180^
C under an inert atmosphere for 2.5 hours.

The reaction mixture is cooled to ambient conditions and

10 600 ml Hcl is added to the mixture which is stirred for 15

minutes. The pH of the mixture is adjusted to pH 3 . Next, 200
ml of water is added into the mixture and stirred for 5 minutes
The mixture is transferred into a 5 L separatory funnel and
extracted three times with 500-ml portions of hexane .

15 The aqueous layer is drained and the combined hexane

solution extracted with four 250-ml portions of 5% NaCl
solut ion .

The hexane is washed 3 times with water. The hexane is
transferred to a flask and the moisture in the hexane removed

20 with anhydrous sodium sulfate (Na^ SOM . The hexane is

filtered through Whatman paper into a clean 1000 ml round
bottom flask and the hexane removed under vacuum with a
rotoevaporator to obtain the CLA. The CLA is stored in a dark
bottle under argon at -80° C until time of use.

25 This method can be modified so as to utilize only food-

grade reagents and solvents as listed in Food Chemicals Codex,
fourth edition. Institute of Medicine, National Academy Press,
1996 .

The active forms of CLA include, in addition to the free
30 acids, the non-toxic salts thereof, the active esters thereof,
such as triglycerides, and mixtures thereof.

The free conjugated linoleic acids (CLA) have been
previously isolated from fried meats and described as
anticarcinogens by Y. L. Ha, N. K. Grimm and M. W, Pariza, in
35 Carcinogenesis, Vol. 8, No. 12, pp. 1881-1887 (1987). Since
then, they have been found in some processed cheese products.
Y. L. Ha, N. K. Grimm and M. W. Pariza, in J. Agric. Food
Che/n., Vol. 37, No. 1, pp. 75-81 (1987). The free acid

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PCTAJS97/04538

-4

forms of the CLA may be prepared by isomerizing linoleic acid
The non-toxic salts of the free acids may be made by reacting
the free acids with a non-toxic base.

One method of synthesizing CLA is described in Example 1.
5 However, CLA may also be prepared from linoleic acid by the
action of a linoleic acid isomerase from a harmless
microorganism, such as the Rumen bacterium Butvrivi hri o
f ibrisolvens . Harmless microorganisms in the intestinal tracts
of rats and other monogastric animals may also convert linoleic

10 acid to CLA (S. F. Chin, J. M. Storkson, W. Liu, K. Allbright
and M. W. Pariza, 1994, J, Wutr. 124; 694-701.

The CLA obtained by the practice of the described methods
of preparation contains one or more of the 9 , 11 -octadecadienoic
acids and/or 10 , 12 -octadecadienoic acids and active isomers

15 thereof. It may be free or bound chemically through ester
linkages. The CLA is heat stable and can be used as is, or
dried and powdered. The CLA is readily con- verted into a non-
toxic salt, such as the sodium or potas-sium salt, by reacting
chemically equivalent amounts of the free acid with an alkali

20 hydroxide at a pH of about 8 to 9. CLA also can be esterified
to glycerol to form mono-, di-, and triglycerides.

Theoretically, 8 possible geometric isomers of 9,11- and
10, 12-octadecadienoic acid (c9, cli; c9,cll; t9,cll; t9,2tll;
cl0,cl2; cl0,tl2; tl0,cl2 and tl0,tl2) would form from the

25 isomerization of c9 , cl2 -octadecadienoic acid. As a result of
the isomerization, only four isomers (c9, oil; c9,tll; tl0,cl2;
and cl0,cl2) would be expected. However, of the four isomers,
c9,tll- and tlO,cl2- isomers are predominantly produced during
the autoxidation or alkali - isomerization of c9, cl2-linoleic

30 acid due to the co-planar characteristics of 5 carbon atoms
around a conjugated double-bond and spatial conflict of the
resonance radical. The remaining two c,c- isomers are minor
contributors ,

The relatively higher distribution of the t,t- isomers of
35 9,11- or 10, 12-octadecadienoic acid apparently results from the
further stabilization of c9,tll- or tl0,cl2- geometric isomers,
which is thermodynamically preferred, during an extended
processing time or long aging period. Additionally the

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PCTaJS97/04538

t,t-isomer of 9,11- or 10 , 12 -octadecadienoic acid that was
predominantly formed during the isomeri zat ion of linoleic
acid geometrical isomers (t9,tl2-, c9,tl2- and t9,cl2-
octadecadienoic acid) may influence the final ratio of the
5 isomers or the final CLA content in the samples.

Liinoleic acid geometrical isomers also influence the
distribution of minor contributors (c,c-isomers of 9,11- and

10.12- , t9,cll- and cll , t 12 -octadecadienoic acids). The

11. 13 - isomer might be produced as a minor product from

10 c9 , cl2 -octadecadienoic acid or from its isomeric forms during
processing .

The exact amount of CLA to be administered to a human to
maintain a level of body fat, of course, can depend upon the
food the human consumes, the form of CLA employed, and the

15 route of administration. It also can depend upon the isomer

ratios. However, generally the amount administered will be the
equivalent of about 1% to about 30% of the weight of the fat in
the human's diet.

The CLA can be administered in food or as pharmaceutical

20 compositions containing the CLA as a free acid; a salt thereof;
an ester thereof, such as a triglyceride; or mixtures thereof.

The amount of CLA to be administered can be expressed as
the amount of CLA based on the total calories consumed daily
by the patient e.g. 0.03 to 3 gram CLA per 100 calories.

25 Alternatively, the amount of CLA can be expressed as a

percentage of the lipid or fat in the food, such as 0.3% to
100% of the food lipid, or as an amount of CLA per gram of food
lipid, such as 3 to 1000 mg. CLA per gram of lipid consumed by
the patient.

30 Generally, the amount of CLA to be administered in

pharmaceutical dosage form will normally be about 100 mg , to
about 20,000 mg. of CLA in the form of the free acids per day.
However, the upper limit of the amount to be employed is not
critical because CLA is relatively non-toxic.

35 The CLA and its non-toxic derivatives, such as the

non- toxic salts, in addition to being added to an animal's food
can be administered in the form of pharmaceutical compositions.

10

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such as tablets, wafer, capsules, solutions or emulsions to the
humans .

The preferred pharmaceutical compositions of CLA contain
the non-toxic sodium, potassium or calcium salt of CLA in
combination with a pharmaceutical diluent. When the
compositions are solutions or suspensions intended for oral
administration, the diluent will be one or more liquid
diluents. When the product is a tablet or capsule, other
conventional diluents, such as lactose, can be employed.

Examples 2 to 4 describe representative foods containing
added CLA.

Example 2

A liquid preparation for parenteral administration zo
humans contains emulsified fat particles of about 0.33-0.5 ^Tr^

15 in diameter. In addition, the emulsions can contain Water for
Injection USP as a diluent, egg phosphatides (1-2%) as an
emulsifying agent and glycerin (2-3%) to adjust toxicity.
These emulsions can be infused intravenously to patients
requiring parenteral nutrition. A preparation for use m the

20 present invention would contain the same ingredients plus 0.5
mg/gm to 10 mg/gm of CLA or alternatively. 0.3% to 100% CLA
based on the food lipid or 0.03 gram to 3 gram per 100 calorie
serving. For such parenteral foods the CLA usually should be
present in the form of the triglycerides.

Example 3

A dietetic margarine for use in the present invention is a
semi -solid or solid vegetable oil -based margarine which, in
addition to the usual ingredients, contains CLA. Such a
margarine will contain about 0.25 mg/gram to about 800 mg/gm of
CLA or about 0.003 gram to 9 gm CLA per 100 calorie serving.

Example 4

A low residue liquid enteral dietetic product useful as a
high-protein, vitamin and mineral supplement contains added
CLA. The amount of CLA present can be about 0.05% to about 5%
35 by weight of the product or about 0.3% to about 100% of the
lipid present or about 0.03 to 3 gram CLA per lOO calories.

30

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-7-

One serving (140 calories) of a representative formula can

contain the following:

Protein (egg white solids) ^ . 5 g

Fat (CLA) 0.1 g

5 Carbohydrate (sucrose, 27. 3g

hydrolyzed corn starch)

Water 1 . 9 g

Vitamins and Minerals (RDA amounts)

It will be readily apparent to those skilled in the art

10 that many other foods, including those described in U.S. Patent

Nos . 4,282,265 and 5,470,839, can be prepared by adding CLA to

the food or by replacing some of the fat in the food with CLA.

The following examples illustrate the practice of the

method of the present invention.

15 Example 5

The level of body fat of a 168 pound (4.75 kg), healthy
human male, age 40 was determined using the "pinch test" on his
waist, thighs and upper arms and his weight was determined by
weighing on a scale. He then was administered four capsules

20 (2400 mg of CLA as the fatty acids) daily and permitted to
consume an unrestricted diet. After eight weeks it was
determined that his weight (4.66 kg) and body fat level had
stabilized at lower levels on an unrestricted diet. CLA
consumption was stopped for one week while food consumption was

25 unrestricted. After the one week period he was weighed and it
was found that three pounds (84,75 g) of body weight had been
gained. The administration of CLA at the original dosage was
resumed for seven weeks whereupon the body weight and body fat
levels returned to the lower levels previously reached after

30 the initial eight weeks of administration of CLA. In the past,
he had normally gained weight and body fat on an unrestricted
diet. Similar results were obtained in several other humans.

Example 6

A healthy 210 pound (5.93 kg) male human, age 53, consumed
35 1200 mg CLA per day for three weeks. During this time his

appetite was somewhat diminished. He then increased his CLA

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-8-

intake to 2400 mg per day and noted a further decrease in
appetite. Throughout this time he did not lose body weight but
noted a decrease in body fat as evidenced by the "pinch" test.

Example 7

5 A healthy 176 pound (4.97 kg) female, age 53, consumed

1200 mg CLA per day. Within three weeks she had lost four
pounds (113 g) , her waistline had decreased by about 1.5 - 2
inches (3.8-5.1 cm), and her appetite had diminished. She
continued taking CLA at the same dose level for three more
10 weeks during which time her body weight and waistline remained
stabilized.

It also will be readily apparent to those skilled in
the art that a number of modifications or changes may be made
without departing from the spirit and scope of the present
15 invention. Therefore, the invention is only to be limited
by the claims.

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rCT/US97/04538

-9-
CLAIMS

1. A mechod of maintaining an existing level of body
fat or body weight of a human desiring to maintain that level
comprises administering to said human a safe and effective
amount of CLA to maintain said existing level.

2. A method of Claim 1 in which the CLA is administered
in a food containing added CLA,

3. A method of Claim 1 in which the CLA is 9,11-
octadecadienoic acid.

4. A method of Claim 1 in which the CLA is 10,12-
10 octadienoic acid.

5. A method of Claim 1 in which the CLA is in the form
of an ester of a conjugated linoleic acid.

6. A method of Claim 1 in which the CLA is in the form
of a non-toxic salt of a conjugated linoleic acid.

15

7. A method of Claim 1 in which the amount of CLA
administered is equal to about 100 mg to about 20,000 mg
per day of the free conjugated linoleic acid.

INTERNATlONAi SEARCH REPORT

Inici jnil ApplicaQon No

PCT/US 97/04538

A. tXASSIHCATION OF SUBJECT MATTER

IPC 6 A61K31/20

According to InicmtticmAl Patent Cla^roricioon (IPC) or to both naoonal cUuficiOon and IPC

B. FIELDS SKARCHED

Minimum documrntioon lurched (classificaoon sy^cm lollowcd by ctaxoficaoon symbolt)

IPC 6 A61K A23D

UocumcnUQcn searched other thin minimum doaimenuooo to ih€ ertcnt that juch documcnu arc included in the field? searched

Electronic data ba.« consulted dunnf the tnicrriauonai search (name of dau ba,«c and, where pracncaJ. :ccarch icrrm uted)

C. DOCUMENTS CONSIDERED TO BE RELEVANT

Category * Ciuoon of document, with indication, where appropriate, of the relevant pasagcs

Relevant to claim No.

FASEB J.,

vol. 10, no. 3, 8 March 1996,
page A56Q XP002034492

PARIZA ET AL.: "Conjugated linoleic acid
(da) reduces body fat"
see abstract nr 3227

EP 0 779 033 A (UNILEVER) 18 June 1997
see page 2 lines 20-22, lines 35-40
see page 5, line 43 - 1 ine 45
see claim 3

-/--

1,2

1-6

Further documcnu are tisted tn tfic conomiaoon of box C.

0

Patent (imily members arc luted in ^nrtex.

' Spcaal categoncs of atcd documents :

'A' document defining the general suu of the an which is not
conadcred to be of particular retcvancc

'E* cariicr document btK published on or afler the intcmabonal
rUing daU

'L* docisncnc which may throw tfoubu on pnonty datn<s) or
which IS QUd to csubluh the publication date of another
aUQon or other spcaal reason (« fpcafied)

*0' document refcmng lo an oral dudosurc. u«, cxhibibon or
other means

*P' doctmcRt published pnor to the intemaao«ul filing tlate but
later than the pnonty dau daimed

T later document publiihed afUr the tntemabonal filing dau
or pnonty daU and not in conflict vath the application but
ated to underttand the pnnapic or theory underlying the
invcnbon

'X' doctment of particular relevance; the datmcd tnvenbon
cannot be considered novel or cannot be considered lo
mvolvc an invcnove step when the document is taken alone

'Y* document of particular relevance; the daimed inventicm
cannot be consdcred to mvolve an inventive step when the
document is combined with one or more other such docu-
ments, such oombicutton being obvious to a person stulled
tn the aft.

'Sl' document member of (he same patent family

Dau of the actual complelion of the inienuoonal uarch

4 July 1997

Dau of mailing of the inicmabonal search report

2 1. 07. 97

NariK and maihng adtat of the ISA

European Patent OfTtce, P.B. Stit PaUndaan 2
NL • 7U0 HV Riiswtik
Td. ( * ]l -70) 340- W40. Til 31 6SI epo nl.
Fac( t 11 70) 340-3016

Aulhontcd officer

Gac, G

Fncm fCTISA'jIt |»ccm4 rfcavt) (>uly iff J)

nAOP 1 nf ?

rNTERNATIONAL SEARCH REPORT

Inter. ful ApptiCAQon No

PCT/US 97/04538

r.(CooonuiQon) DOCUMENTS CONSIDERED TO BE RELEVANT

Clat£jory *

CiUttoo ol docimicni, with indicaoon, w^cre »pprT>pn*te. of Uk rclcvuil p&uiies f

Iclcvwt lo claim No.

BIOCHEM. 8I0PHYS. RES. COMMUN..
vol. 198, no. 3. 15 February 1994,
pages 1107-1112. XPOG0567057
MILLER ET AL. : "Feeding Conjugated
linoleic acid to animals partially
overcomes catabolic responses due to
endotoxin injection"
see the whole document

EP 0 579 901 A (WISCONSIN ALUMINI RESEARCH
FOUNDATION) 26 January 1994
see the whole document

WO 96 06605 A (WISCONSIN ALUMINI RESEARCH
FOUNDATION) 7 March 1996
see whole document, especially page 6
lines 1-4, page 7, lines 12-22, page 10
lines 5-16

WO 94 16690 A (WISCONSIN ALUMINI RESEARCH
FOUNDATION) 4 August 1994
see the whole document

JOURNAL OF NUTRITION.

vol. 124, no. 5, 1 January 1994,

pages 694-701, XPOO0574585

CHIN S F ET AL: "CONJUGATED LINOLEIC ACID

(9,11- AND 10,12-OCTADECAOIENOIC ACID) IS

PRODUCED IN CONVENTIONAL BUT NOT GERM-FREE

RATS FED LINOLEIC ACID"

see page 696 left column second line.

right column "results experiment 1"

second paragraph

see Figures 3A and 38

1,2

1.2

1-7

1-7

1-4

1

Form PC T.I S A.'} 10 (CBntmucuon of MODn^ ihMt) (July Iff] I

INTERNATIONAL SEARCH REPORT

naiion&l Applicaxion No

PCT/US 97/04S38

Box I Observations where certain daims were found unsearchable (Coniinuaiion of item I of first sheet)

This Intcrnauonal Search Report hu not been esubltshcd in respect of ceruin cizims under Articte 17(3X0 the following reuons:
1. [T] Claims Noj.:

because they leiaie lo subfcct matter not required to be searched by this Authority, namely:

Remark: Although claim(s) 1-7

Is(are) directed to a method of treatment of the human/animal
body, the search has been carried out and based on the alleged
effects of the compound/composition.

7. \ ] Claims Noj.:

because they relate to parts of the Inter nauonal ApplKaiion thai do not comply wuh the prescribed requtrcmenu to such
an extent thai no meaningful Intcrnauonal Search can be carried out. specifically:

I I Claims Nos.;

because they are dependent claims and are not dra/ied in accordance wiih the second and third sentences of Rule 6.4(a).

Box II Observations where unity of invention is lacking <Continumtion of item 2 of first sheet)

This Iniernauonal Searching Authoruy found multiple mvenuons in this iniernauonal appltcauon. as follows:

1 As aJI required additional search fees were umely paid by the applicant, this ImernationaJ Search Report covers all
* — ' searchable dajms.

2 CD searchable claims could be searched without effort N*ufying an addiuonal fee. this Authority did not invite payment

of any additional fee.

3. I I "me of the required additional search fees were umely paid by the applicant, this ImcrnauonaJ Search Report

I— I covers only those daixm for which feei were paid, specifically daims Nos.i

4. Q No required addiuonaJ search fees were umely paid by the applicant ConsequcnUy. this Inurnauonal Search Report is
restricted to the invention first mcnuoncd in the dums. it is covered by claims Nos.,

Remark oa Pratest j~j jhe additional search fees %w:re accompanied by the applicant s prousL

I I No protest accompanied the payment of addiuonal search fees.

Form PCT.'1SA.2I0 (continuation of first sheet (l))(July 1993)

INTERNATIONAL SEAJICH REPORT

Infonnaoon on patent (imily members

Inlc. .vnal Applicjaoo No

PCT/US 97/04538

Paicnt document

Publication

Patent family

PubltcaLion

cited in search report

dale

m€mber(j)

dau

lA HA 07

All

AU

7175496

A

T) nc rtT

22-05-97

LA

2190241

A

15-05-97

EP 579901 A

26-01-94

US

5430066 A

04-07-95

DE

69301693

0

11-04-96

DE

69301693

T

25-07-96

US

5428072

A

27-06-95

lie

US

5504114

A

02-04-95

lie

5554645

A

10-09-96

A7 nc

1 IC

us

5554646

A

10-09-96

EP

0731699

A

18-09-96

WO 9416690 A

04-08-94

US

5428072

A

27-06-95

EP

0680318

A

08-11-95

JP

8505775

T

25-06-96

US

5504114

A

02-04-96

US

5554646 A

10-09-96

Form PCT.1SA/]tt (^twii family winci) |/«^ lt«])